小麦花粉致死基因Ki的SSR标记分析

小麦雄性不育主要是通过花粉的败育表现,其不育材料对小麦杂种优势的利用研究具有重要意义和价值,国外研究表明,某些特定普通小麦品种间杂交F₁表现的花粉部分不育现象,受控于核基因组花粉致死基因Ki,为了筛选小麦花粉致死基因Ki的连锁标记,利用现代分子生物学技术通过定位该基因,克隆出花粉致死基因连锁标记片段,为小麦雄性不育种质材料的转育提供有效的选择标记。对小麦花粉致死基因Ki进行了分子标记定位,以‘中国春’和澳大利亚春小麦品种的BC₁F₁代作为定位群体,利用分离群体分组分析法(BSA)对位于小麦6B染色体上85对SSR引物进行多态性筛选,具有多态性的引物再通过BC₁F₁定位群体进行验证,从中筛选出与目的基因连锁的2个SSR标记Xgwm626和Xgpw4138。运用Mapmaker 3.0软件进行连锁分析。结果表明,Xgwm626和Xgpw4138与Ki基因的遗传距离分别为9.2 cM和6.9 cM,且2个SSR标记位于目的基因两侧,并将Ki定位于小麦6BL染色体上。研究结果为Ki基因的分子标记辅助选择和进一步精细定位奠定了基础。     

水稻‘陆18S’核不育基因连锁的分子标记

为了研究水稻不育基因的遗传机理和连锁情况,利用‘陆18S’与3个父本杂交,并对F2进行花粉育性和结实率观察和遗传分析,同时对F2后代进行RAPD分子标记。通过遗传分析得知,‘陆18S’不育系由1对隐性核基因控制。通过分子标记的筛选,找到了与‘陆18S’雄性不育紧密连锁的RAPD标记S490,片段大小为850 bp。获得与不育基因紧密连锁的RAPD标记。为不育基因克隆、定位奠定了坚实的基础。     

水稻特异亲和基因S-e的分子定位

水稻籼粳亚种间杂种具有强大的优势，但亚种间杂种的不育性限制了这一优势的利用。开展杂种不育基因的定位工作，对于进一步了解杂种不育性的遗传基础，克服亚种间杂种的不育性具有重要的意义。本研究选用粳型品种台中65的近等基因系E47-1和籼型品种广陆矮4号为材料，利用74个SSR标记对杂种F2群体进行偏态分离标记的筛选，同时根据F2和F3群体花粉育性和具有偏态分离的SSR标记之间的连锁关系，对特异亲和基因(F1花粉不育基因)S-e座位进行了分子定位，取得了以下主要结果：1、利用116个均匀分布在水稻12条染色体上的SSR标记对籼粳两亲本进行多态性筛选。结果有101个SSR标记在亲本间具有多态性，15个SSR标记在亲本间无多态性，SSR标记在亲本间的多态率高达87．07％。2、选用74个亲本间具有多态性的SSR标记对E47-1／广陆矮4号组合F2群体的偏态分离进行了初步的筛选和分析。发现有6个染色体区段的9个SSR标记在F2群体中存在偏态分离，它们分别位于第3、第6、第7、第10、第11和第12染色体上，卡方值均达到显著或极显著水平。6个染色体区段中有2个严重偏态分离区段，分别位于第6和第12染色体。3、通过对F2群体的花粉育性和偏态分离区段的SSR标记基因型的相关关系分析，表明位于第12染色体上的SSR标记RMl9附近存在一个F1花粉不育基因。继而在该标记附近设计位置特异性微卫星标记PSM401、PSMl80、PSMl82，利用F3作图群体，将特异亲和基因S-e座位定位在分子标记PSM401、PSMl80和PSMl82、RMl9之间，该基因与各标记的遗传距离分别为2．3cM、1．3cM、3．7cM和4．3cM。4、选取在S-a、s-b、S-c、S-d、S-e五个座位均纯合、花粉表现为部分不育的F2单株，发展了另一R群体，表明该群体存在另一特异亲和基因座s-f。本研究利用SSR标记，对特异亲和基因S-e进行了分子定位。S-e座位的分子定位，进一步丰富和完善了特异亲和性的学术观点，并为分子标记辅助选育水稻的粳型亲籼系奠定了基础。     

陆地棉光敏雄性不育基因ys-1的遗传分析与初步定位

【目的】精细定位和克隆陆地棉光敏雄性不育基因。【方法】从陆地棉中040029的航天诱变后代中选育出新型光敏雄性不育系中9106,以中9106与11个陆地棉材料配制杂交组合,初步分析了中9106的杂种优势。以中9106为母本与陆地棉乐土603构建遗传分离群体F1、F2,分析不育性状的遗传特征。利用集团分离分析法筛选简单序列重复(Simple sequence repeat,SSR),并在包含186个单株的中9106×乐土603 F2群体中用上述SSR初步定位不育基因。【结果】中9106为母本×陆地棉乐土603的F1单株育性均表现正常,而F2群体中的可育单株数∶不育单株数符合3∶1的分离比,推测中9106的不育性状受1对隐性核基因控制。利用集团分离分析法从16 544对SSR中筛选到18对与该不育基因连锁的标记。利用中9106×乐土603的F2群体和18对SSR标记,将不育基因定位于D12染色体,位于标记NAU3442与CGR6339之间,遗传距离均为0.2c M,将该不育基因命名为ys-1。【结论】本研究有助于ys-1的精细定位及其克隆。     

谷子6号染色体雄性不育基因的克隆

克隆谷子雄性不育材料1066A的不育基因,分析不育基因与可育基因存在的突变位点,为揭示谷子雄性不育分子机制、利用分子标记辅助选择方法选育多用途的不育材料奠定基础。利用谷子全基因组测序数据及前人不育基因定位结果克隆谷子雄性不育材料1066A的雄性不育基因,发掘导致不育的突变位点,旨在为从分子水平揭示谷子不育机制、利用分子标记辅助选择方法选育多用途的不育材料奠定基础。首先利用生物信息学方法从豫谷1号6号染色体找到1个雄性不育基因位点(Si015780m.g),该基因全长5 027个碱基,编码479个氨基酸,且位于前人用分子标记定位的基因组区间内。根据豫谷1号不育基因序列设计2对特异引物在雄性不育材料1066A的基因组DNA进行PCR扩增。将扩增产生的2个基因组片段进行拼接后在谷子不育材料1066A中获得2 561 bp的基因序列,包含了下游部分编码区。通过对豫谷1号、张谷、1066A的不育基因部分编码序列及推定的蛋白质序列进行比对分析,结果发现谷子不育材料1066A的不育基因编码序列存在3处突变:2处单碱基替换和1处单碱基插入,这3处突变导致谷子不育材料1066A的不育基因蛋白的第402,403个氨基酸由异亮氨酸和亮氨酸替换成缬氨酸和异亮氨酸,同时导致其不育基因编码的蛋白在第466个氨基酸处发生提前终止。3处突变中2处氨基酸替换对编码蛋白的功能影响不大,因此,认为谷子不育材料1066A的不育基因蛋白翻译提前终止可能是导致其产生不育的原因。     

水稻F1花粉不育基因的精细定位及其遗传分化研究

水稻籼粳亚种间杂种的不育性限制了亚种间的遗传交流和杂种优势利用。本研究通过发展位置特异性的微卫星标记将F1花粉不育基因S-6座位进行了精细定位；通过分析近等基因系中代换片段的遗传效应，鉴定出了F1花粉不育基因S-d座位，利用位置特异性的微卫星标记将S-d进行了定位；根据基因组的序列资料和利用较大的作图群体对S-6和S-d两个座位进行了物理作图；通过分子标记辅助选择培育了一批复等位基因近等基因系，对育性基因的遗传分化进行了研究。取得了如下主要结果：1、根据S-6座位初步定位的结果发展位置特异性的微卫星标记，将F1花粉不育基因座S-6进行了精细定位。结果表明多态性标记均与S-6座位紧密连锁，其中R830STS、PSM7、PSM8、PSM9、．PSM59和PSM60位于S-6座位一端，与S-6座位遗传距离分别为1．5cM、1．2cM、0．9cM、0．9cM、0．9cM和0．9cM，而PSM202、PSM206、PSM208、RMl3、R2213SSTS和RM413位于S-6座位的另一端，与S-6座位的遗传距离分别为0．9cM、2．1cM、3．8cM、4．1cM、4．4cM和5．3cM。2、根据S-6座位精细定位的结果，从IRGSP下载了S-6座位所在区域克隆的序列，将克隆的序列进行了拼接，同时将与S-6座位紧密连锁的分子标记与序列拼接图进行了电子整合。根据整合的结果发展位置特异性的微卫星标记和STS标记，利用500株的作图群体，最终将S-6座位界定在PSM8与PSM215之间182．2kb的范围，其中PSM214、T17、T18和T19与S-6座位完全连锁。3、通过对近等基因系E11-5中代换片段遗传效应的分析，在第1染色体的代换片段上鉴定出一个新的F1花粉不育基因座S-d。根据基因组的序列发展位置特异性的微卫星标记将S-d座位进行了定位。结果表明多态性标记均与S-d座位紧密连锁，其中PSM27、PSM24、．PSM26、PSM23、PSM31、PSM25、PSM37、PSM41、PSM42、PSM43、．PSM44、．PSMl2和PSMl3位于S-d座位的一端，与S-d座位的遗传距离分别为10．6cM、7．2cM、7．2cM、6．8cM、6．8cM、6．4cM、6．4cM、4．8cM、4．8cM、3．2cM、1．6cM、0．4cM和0．4cM，而RM84、RM86、RM323、RMl和RM283位于S-d座位的另一端，与S-d座位的遗传距离分别为3．8cM、4．6cM、6．7cM、7．5cM和8．7cM。4、根据S—d座位定位的结果，从IRGSP下载了S-d座位所在区域克隆的序列，将克隆的序列进行了拼接，同时将与S-d紧密连锁的分子标记与序列拼接图进行了电子整合。根据整合的结果发展位置特异性的微卫星标记和STS标记，利用2160株的作图群体，最终将S-d座位界定在67．8kb的范围内，其中PSM93和PSM74位于S-d座位的两侧且各与S-d仅有一个重组，而PSM95、PSM96、T1和T2与S-d座位完全连锁。RiceGAAS注释分析表明在此区段有17个ORF。，其中3个ORF可能与杂种不育性有关。BLAST分析表明此段序列籼粳之间的同源性较低，这也可能是杂种不育的一个原因。5、通过分子标记辅助选择，在F1花粉不育基因s-n、s-6、s-c和s-d座位各培育了一批复等位基因近等基因系，测交分析表明各不育基因座位上的不育基因不仅分化为相对的S^i和S^j，而且基因型类型相同的复等位基因的遗传分化也达到了显著差异的水平。携带有多个复等位基因的近等基因系的测交分析表明，复等位基因近等基因系的遗传效应为各基因座位遗传效应的累加，各基因的遗传效应相互独立，彼此间无相互作用。     

水稻S-c座位的PCR标记精细定位及分子标记辅助选择

以粳型品种台中65及其近等基因F1不育系TISL5为材料,利用STS和SSLP标记对水稻F1花粉不育基因座位S-c进行了精细定位,RG227STS和RM218分别位于S-c的两侧,与S-c的距离分别为0.3cM和4.3cM.通过对40个籼、粳和中间型品系标记基因型的鉴定,分析了分子标记基因型与S-c基因型之间的关系,建立了以PCR为基础的分子标记辅助选择体系.利     

水稻光敏核不育基因pms3的精细定位

为了进一步研究水稻晚粳品种农垦58转变为光敏核不育水稻农垦58S的突变位点,并精细定位光敏不育基因pms3,我们利用农垦58S/1514杂交组合的F1进行花药培养构建了一个DH群体,验证了在该群体中光敏不育受pms1、 pms3两对基因的控制,并根据分子标记分析选择第12染色体是1514基因型、 第7染色体农垦58S基因型的DH系DH80与农垦58S杂     

一个粳稻来源抗稻瘟病基因的鉴定、遗传分析和基因定位

7001S是一个广谱抗稻瘟病的粳稻两用核不育系,对来自全国不同稻区的22株稻瘟病菌系均表现为高度抗性。通过构建7001S/80-4B F2群体的遗传分析和初步定位表明,F2分离单株对稻瘟病菌的抗性呈明显的抗、感双峰分布,抗感分离符合3﹕1的理论比例,说明粳稻7001S对稻瘟病菌的抗性由1对显性核基因或一个显性QTL位点控制,并将该基因初步定位于第11染色体长臂末端。进一步通过扩大遗传群体和分子标记开发,利用基于BSA的隐性群体分析技术,将目的基因精细定位于P21-2415和RM27322之间约310 kb的范围内,并获得了可用于分子标记辅助选择的紧密连锁和共分离分子标记,同时对目标基因所在区域进行基因预测,初步确定了候选基因。为进一步开展该抗稻瘟病基因的克隆、功能验证和抗病机理研究,以及通过分子标记辅助选择技术培育抗稻瘟病水稻新品种等工作奠定了基础。     

水稻雌性不育分子机理研究进展

育性是水稻的关键农艺性状,其发生机理和遗传基础是发育生物学的研究热点。不育可分为雄性不育和雌性不育。雌性不育由于性状难鉴定、突变体难获得及内部机制复杂等原因,其研究远远滞后于雄性不育。近年来,由于水稻雌性不育突变体的不断发现及分子生物学研究手段的快速进步,一批控制水稻雌性不育的基因相继被定位和克隆,为揭示水稻雌性不育发生机理奠定了基础。本文综述了近二十年来水稻雌性不育分子遗传机理的有关研究进展,主要包括杂种不育基因,影响胚珠发育、细胞分裂和花器官发育的基因,以及转录组等方面的研究,以期加深对水稻雌性不育的认识,促进对水稻雌性不育的探索和利用。     

A note on the origin of Kalanchoë x vadensis Boom & Zeilinga (Crassulaceae)

Summary K x vadensis is a hybrid of K. blossfeldiana and K. marmorata obtained after doubling the number of chromosomes.     

Avoidance of leaf rust fungi in wild relatives of cultivated cereals

Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed.     

Relation between wheat-rye crossability and seed set of common wheat after pollination with other species in the Hordeae

Summary Cultivars of common wheat (Triticum aestivum L. em. Thell.) of high wheat-rye (Secale cereale L.) crossability set more seed with pollen of other related species than did wheats of low wheat-rye crossability. This was found to be true for pollen parents from the genera Triticum, Aegilops, Secale, Agropyron and Elymus.     

Mechanisms and diversity of resistance to sorghum shoot fly,Atherigona soccata

Sorghum shoot fly, Atherigona soccata, is one of the important pests of postrainy season sorghums. Of the 90 sorghum genotypes evaluated for resistance to this pest, RHRB 12, ICSV 713, 25026, 93046 and 25027, IS 33844‐5, Giddi Maldandi and RVRT 3 exhibited resistance in postrainy season, while ICSB 463, Phule Anuradha, RHRB 19, Parbhani Moti, ICSV 705, PS 35805, IS 5480, 5622, 17726, 18368 and 34722, RVRT 1, ICSR 93031 and Dagidi Solapur showed resistance in rainy season, suggesting season‐specific expression of resistance to A. soccata. ICSB 461, ICSB 463, Phule Yasodha, M 35‐1, ICSV 700, 711, 25010, 25019 and 93089, IS 18662, Phule Vasudha, IS 18551 and 33844‐5 and Barsizoot had fewer deadhearts than plants with eggs across seasons, suggesting antibiosis as one of the resistance mechanism. Five genotypes exhibited resistance with high grain yield across seasons. Correlation, path and stepwise regression analyses indicated that leaf glossiness, seedling vigour, trichome density, oviposition and leaf sheath pigmentation were associated with the expression of resistance/susceptibility to shoot fly, and these can be used as marker traits to select and develop shoot fly‐resistant sorghums.     

Reaction of tritordeum to Fusarium culmorum and Septoria nodorum

Summary Hordeum chilense is a wild barley extensively used in wide crosses in the Triticeae. It could be a valuable source of resistance to Fusarium culmorum and Septoria nodorum. Some H. chilense x Triticum spp. amphiploids, named tritordeums, were more resistant than the parental wheat line to these diseases, others were not. Average contents of ergosterol and deoxynivalenol (DON) suggested that resistance to colonization by Fusarium was the highest for Hordeum chilense, followed by tritordeum and wheat in decreasing order. In particular, the H. chilense genotypes H7 and H17 enhanced the wheat resistance to F. culmorum in its tritordeum offsprings. Resistance to S. nodorum in tritordeum was not associated with tall plant height. There is sufficient genetic variation for resistance to F. culmorum and S. nodorum among tritordeum to allow the breeding of lines combining short straw and resistance to both diseases.     

Breeding apomictic bahiagrass (Paspalum notatum Flügge) with improved turf traits

The development of a turf‐type bahiagrass could have a remarkable impact on the billion‐dollar turf industry in the south‐eastern United States. The goals of this study were to (i) select bahiagrass genotypes with improved turf attributes and (ii) determine the reproductive mode and seed fertility of selected genotypes. The population included mutants, wild types, hybrids and standard cultivars evaluated at two locations for two years. Morphological variation was observed and clonal repeatability (H²) ranged from 0.27 to 0.90. Flowering was extremely reduced in some mutants, which also exhibited dark green colour, finer texture and reduced canopy height. Cytoembryological observations revealed that diploids reproduced sexually and tetraploids by obligate or facultative apomixis. Genotypes with superior seed fertility were identified, while others were comparable to the standard cultivars. While mutant plants exhibited great morphological variation, the random mutations did not affect their reproductive mode, validating the use of mutagenesis in apomictic species. The high H² indicated that selection for improved turf attributes could be successfully performed with the advantage of clonal seed propagation.     

Disease resistance in protected crops and mushrooms

Summary Cultivars of tomatoes, cucumbers, lettuce and peppers have been bred for resistance to one or more pathogens. Some tomato and cucumber cultivars have resistance to a wide range of diseases. Resistance has been transient in many cases and a succession of cultivars with new genes or new combinations of resistance genes has been necessary to maintain control. There has been a number of notable exceptions and these have included durable resistance to such pathogens asFulvia fulva and tomato mosaic virus. With lettuce the resistance situation is complicated by the occurrence of fungicide resistant pathotypes. There are no strains ofAgaricus bisporus purposely bred for disease resistance.In protected flower crops only resistance to Fusarium wilt in carnations has been purposely bred but differences in disease resistance are apparent in cultivars of many ornamental crops. This is particularly so in chrysanthemums where there are cultivars with resistance to many of the major pathogens. Similar situations occur with other flower crops and pot plants. Cultivars of some species have not been systematically investigated for resistance.The need for genetic resistance will increase with the further reduction, in the limits on pesticide use and an increasing public awareness and importance of pesticide pollution.ADAS is an executive agency of the Ministry of Agiculture, Fisheries and Food and the Welsh Office.     

Genetic constitution and diversity in four narrow endemic redwoods from the family Cupressaceae

The genetic constitution and diversity of four relictual redwoods are discussed in this review. These include monotypic genera of the family Cupressaceae: coast redwood (Sequoia sempervirens), giant sequoia (Sequoiadendron giganteum), dawn redwood (Metasequoia glyptostroboides), and alerce (Fitzroya cupressoides). All four species are narrow endemics, share a number of common phenotypic traits, including red wood, and are threatened species. Fossil history suggests that the ancestors of redwoods probably originated during the Cretaceous and Tertiary periods and flourished thereafter for millions of years. Towards the end of the Tertiary period began their decline and struggle for existence that continued during the subsequent geologic upheavals and climate changes, until the survival of the present-day redwoods in the current restricted locations in the world (USA, China, and South America). Although two species, Sequoiadendron and Metasequoia, are diploids (2n = 22), and the other two are polyploids: Fitzroya a tetraploid (2n = 4x = 44), and Sequoia a hexaploid (2n = 6x = 66); they all share the same basic chromosome number x = 11. The genome size in the hexaploid Sequoia is one of the largest (31,500 MB) in the conifers, while the genome sizes of diploid Metasequoia and Sequoiadendron are about one-third (~10,000 MB) of Sequoia. Genetic diversity in the redwoods is lower than most other gymnosperms, except in Sequoia, which seems to rank near the upper quarter of the coniferous forest trees. Genomic research is sparse in the redwoods, and should be pursued for a better understanding of their genome structure, function, and adaptive genetic diversity.     

Inheritance of fruit colour in a wild Russian red raspberry seedling

Summary A new gene, i, from a self-incompatible wild Russian Rubus idaeus seedling is described. The gene i interacts with the fruit colour gene T, Tii plants having apricot instead of red fruits, and spines, leaves and stems pigmented to varying degrees. A deficit of apricot-fruited seedlings in progenies segregating for i could be explained by linkage between i and the self-incompatibility S locus with a crossover value of approximately 26.7%.Segregations for spine colour in crosses and backcrosses of the Russian seedling with two green-spined (ttII) raspberries selected for their yellow fruit colour, showed that both the latter carried the spine colour intensifier gene P. This suggests that earlier authors were incorrect in attributing the apricot fruit colour developed by some tt plants to a pleiotropic effect of P.     

The induction in vitro of adventitious shoots in Rosa

Summary Adventitious shoots were formed on excised leaves, roots and callus of Rosa persica x xanthina and on excised leaves of R. laevigata and R. wichuraiana on culture media that included BAP and NAA as growth regulators. Shoots formed freely on freshly cultured callus of R. persica x xanthina but their production declined in successive cultures and ceased after twelve weeks. Transplantation to soil was improved by rooting plantlets in cellulose plugs in vitro and transferring plantlets to soil while still in the plugs.